Efficient design and assembly of custom talens using the golden gate platform

Research output: Chapter in Book/Report/Conference proceedingChapter

27 Scopus citations

Abstract

An important breakthrough in the field of genome engineering was the discovery of the modular T ranscription A ctivator- L ike E ffector (TALE) DNA binding domain and the development of TALE n ucleases (TALENs). TALENs enable researchers to make DNA double-strand breaks in target loci to create gene knockouts or introduce specific DNA sequence modifications. Precise genome engineering is increasingly being used to study gene function, develop disease models or create new traits in crop species. Underlying the boom in genome engineering is the striking simplicity and low cost of engineering new specificities of TALENs and other sequence-specific nucleases. In this chapter, we describe a rapid, inexpensive, and user-friendly protocol for custom TALEN construction based on one of the most popular TALEN assembly platforms, the Golden Gate cloning method. Using this protocol, ready-to-use TALENs with specificity for targets 13-32 bp long are constructed within 5 days.

Original languageEnglish (US)
Title of host publicationChromosomal Mutagenesis
Subtitle of host publicationSecond Edition
PublisherSpringer Fachmedien
Pages133-159
Number of pages27
ISBN (Electronic)9781493918621
ISBN (Print)9781493918614
DOIs
StatePublished - Nov 19 2014

Keywords

  • Genome engineering
  • Golden Gate assembly
  • Site-specific nuclease
  • TALEN
  • Targeted mutagenesis

Fingerprint Dive into the research topics of 'Efficient design and assembly of custom talens using the golden gate platform'. Together they form a unique fingerprint.

Cite this